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<p>Single-molecule localization microscopy achieves nanometer-scale resolution but is compromised by sample drift during image acquisition. Here we present reinforced optical cage systems, a novel approach that eliminates drift at its mechanical source rather than correcting it through complex image post-processing or fiducial markers. Reinforced optical cage systems employ perforated optomechanical components interconnected by tungsten-steel rods in a design proven by mechanical stability simulations. Our bench-top microscope, built with reinforced optical cage systems, demonstrated exceptional three-dimensional stability, with mean cumulative lateral drift of approximately 5 nanometers over 2 h in widefield fluorescence microscopy and 11-16 nanometers over 15 min in single-molecule localization microscopy, free from measurable axial drift. This development allows super-resolution microscopy to reach its full resolution without the necessity of sample drift correction, offering a straightforward, cost-effective, low-maintenance, and readily accessible solution to high-performance super-resolution microscopy. By addressing the fundamental issue of mechanical instability, reinforced optical cage systems enable improved precision instrumentation for the broader scientific and engineering community.</p>

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Reinforced optical cage systems enable drift-free single-molecule localization microscopy

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月15日

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  • Hao Qiu,
  • Matthew C. Tang,
  • Selene K. Roberts,
  • Guoli Li,
  • Rong Su,
  • Marisa L. Martin-Fernandez,
  • David T. Clarke,
  • Shugang Liu,
  • Xiaojie Liu,
  • Lin Wang

摘要

Single-molecule localization microscopy achieves nanometer-scale resolution but is compromised by sample drift during image acquisition. Here we present reinforced optical cage systems, a novel approach that eliminates drift at its mechanical source rather than correcting it through complex image post-processing or fiducial markers. Reinforced optical cage systems employ perforated optomechanical components interconnected by tungsten-steel rods in a design proven by mechanical stability simulations. Our bench-top microscope, built with reinforced optical cage systems, demonstrated exceptional three-dimensional stability, with mean cumulative lateral drift of approximately 5 nanometers over 2 h in widefield fluorescence microscopy and 11-16 nanometers over 15 min in single-molecule localization microscopy, free from measurable axial drift. This development allows super-resolution microscopy to reach its full resolution without the necessity of sample drift correction, offering a straightforward, cost-effective, low-maintenance, and readily accessible solution to high-performance super-resolution microscopy. By addressing the fundamental issue of mechanical instability, reinforced optical cage systems enable improved precision instrumentation for the broader scientific and engineering community.